Using the research article attached, provide an overview of the study and describe the strategy that was used to select the sample from the population. Evaluate the effectiveness of the sampling meth

Contents lists available atScienceDirect Food Quality and Preference journal homepage:www.elsevier.com/locate/foodqual Short Communication Comparison of two sugar reduction strategies with children: Case study with grape nectars Mayara Lima a,⁎ , Gastón Ares b, Rosires Deliza c aFood Technology Department, Technology Institute, Federal Rural University of Rio de Janeiro, Rodovia BR 456, km 7, Seropédica, RJ, BrazilbSensometrics & Consumer Science, Instituto Polo Tecnológico de Pando, Facultad de Química, Universidad de la República, By Pass de Rutas 8 y 101 s/n, Pando, UruguaycEmbrapa Agroindústria de Alimentos, Avenida das Américas, 29501 CEP 23020-470 Rio de Janeiro, RJ, Brazil ARTICLE INFO Keywords:

Children Added-sugar reduction Sugar-sweetened beverages Check-all-that-apply ABSTRACT Excessive sugar consumption has been identi ﬁed as the main dietary determinants of obesity among children and adolescents. Sugar sweetened beverages are one of the main sources of added sugar in the diet. Therefore, gradual sugar reduction in these products can contribute to decreasing sugar intake and encourage children to slowly develop preference for products with lower sugar concentration. However, speci ﬁc recommendations on how to implement gradual sugar reduction programs are still lacking. The aim of the present study was to compare two sugar reduction strategies (stepwise vs. gradual) on children ’s sensory and hedonic perception of grape nectar. One hundred and seventeen 6 –12 years old children participated in a two-part study: a 9-week study and a ﬁnal liking test at the end of 11 weeks. In the 9-week study, children were divided in two sugar reduction strategies: stepwise and gradual. Children ’s sensory and hedonic perception was evaluated using 9- point hedonic scales and check-all-that-apply questions. There were no major di ﬀerences between the gradual and stepwise reduction groups in children ’s sensory and hedonic perception. However, results showed that the gradual reduction strategy led to smaller changes in children ’s sensory and hedonic perception than the stepwise strategy. In the ﬁnal test, children in the stepwise group gave signi ﬁcantly higher liking scores to the samples than those in the gradual sugar reduction strategy. Comparing the overall liking between the ﬁrst time the children tasted the grape nectars and the ﬁnal test, a signi ﬁcant di ﬀerence was observed for the sample with lower sugar content only for the stepwise group, which was higher in the 11th week. Although di ﬀerences between the sugar reduction strategies were small, the gradual reduction strategy seems to be recommended over the step-wise strategy. 1. Introduction Excessive sugar consumption has been identi ﬁed as the major cause of excessive caloric intake and the main dietary determinant of obesity among children and adolescents ( Ambrosini, Johns, Northstone, Emmett, & Jebb, 2016; Te Morenga, Mallard, & Mann, 2012 ). In par- ticular, sugar-sweetened beverages (SSB), including soda, fruit- ﬂavored drinks, and sport drinks, are one of the main sources of sugars in children ’s diets ( Malik, Pan, Willett, & Hu, 2013 ). According to Garriguet (2008) , consumption of sugar from SSB alone accounts for the total recommended daily free sugar intake (10% of the total energy intake; World Health Organization, 2015 ), suggesting that SSB should be a prime target for reducing sugar intake among children. Considering that changes in eating habits are di ﬃcult to achieve, strategies that do not require consumers ’willpower to change have the greatest chance of succeeding at the population level in the short term ( Dobbs et al., 2014). For this reason, reformulation of products has been proposed as one of the most e ﬀective strategies to encourage changes in nutrient intake ( van Raaij, Hendriksen, & Verhagen, 2009). If the added sugar content of products is reduced without consumers ’awareness, it can lead to a reduction in their sugar intake even if they do not change their eating behavior ( MacGregor & Hashem, 2014).

The main challenge for reducing the added sugar content of food products, particularly in the case of sweetened beverages, is that it causes changes in their sensory characteristics, which are key de- terminants of consumers ’liking ( van Raaij et al., 2009). This is parti- cularly relevant in the case of children due to their heightened pre- ference for sweet taste ( Ventura & Menella, 2011 ).

There are two main strategies for sugar reduction without sub- stitution: abrupt sugar reduction, which consists of reducing sugar in a single step, and gradual sugar reduction, which consists of consecutive small reductions. Gradual sugar reduction has been shown to be more https://doi.org/10.1016/j.foodqual.2018.07.002 Received 26 April 2018; Received in revised form 29 June 2018; Accepted 2 July 2018 ⁎Corresponding author.

E-mail address: [email protected] (M. Lima). Food Quality and Preference 71 (2019) 163–167 Available online 02 July 2018 0950-3293/ © 2018 Elsevier Ltd. All rights reserved. T eﬀ ective than the abrupt sugar reduction, as consumers do not notice changes in the sensory characteristics of the products, and they slowly become accustomed to lower sugar concentration ( MacGregor & Hashem, 2014 ). However, speci ﬁc recommendations on how to im- plement gradual sugar reduction programs are still lacking. Reductions lower than the di ﬀerence threshold for added sugar have been re- commended as they assure that the great majority of the consumers would not notice the change ( MacGregor & Hashem, 2014). However, this could entail several reduction steps to achieve the sugar reduction target, which would not be practical for food companies, and would require long implementation times. Therefore, a stepwise sugar reduc- tion strategy based on larger reduction steps would provide practical advantages over gradual reduction programs based on di ﬀerence thresholds. However, this strategy could lead to larger changes in consumers ’sensory and hedonic product perception.

In this context, the aim of the present study was to compare two sugar reduction strategies (stepwise vs. gradual) on children ’s sensory and hedonic perception of grape nectar. It was hypothesized that the implementation of cumulative unnoticeable reductions in the added sugar content of the nectars would have a smaller impact on children ’s sensory and hedonic perception than stepwise reductions.

2. Materials and methods 2.1. Participants A total of 147 children, aged 6– 12 years old (46% female), were recruited in a private elementary school in Rio de Janeiro (Brazil).

Children and their parents signed informed consent forms to participate in the study. Ethical approval was obtained from the ethics committee of Universidade Federal do Estado do Rio de Janeiro (Plataforma Brasil CAAE: 55023416.0.0000.5285). Children received a small gift for their participation in each session.

2.2. Nectar formulation Grape nectar samples were formulated by diluting a commercial grape juice with no added sugar (Del Valle, Brazil) to 35% using mi- neral water (Minalba, Brazil). The dilution of grape juice was de- termined based on the characteristics of commercial products available in the marketplace and pilot testing. The nectars were added with 4.3 –10% commercial sugar (União, Brazil). The ingredients were weighed in a semi analytical balance and manually mixed. The nectars were stored under refrigeration for one day before being served to children.

The initial added-sugar concentration of the nectars (10%) was se- lected based on commercial grape nectars available in the Brazilian marketplace, whereas the lowest concentration (4.3%) was expected to cause a signi ﬁcant decrease in children ’s hedonic perception ( Lima, Ares, & Deliza, 2018 ). The added-sugar concentration of the nectars was reduced according to two strategies: stepwise and gradual ( Table 1). In the gradual strategy, added-sugar content was reduced from 10% to 4.3% according to the di ﬀerence thresholds for added sugar estimated in a previous study ( Lima et al., 2018). In the stepwise strategy, added- sugar content was reduced from 10.0 to 4.3 in two steps ( Table 1).

2.3. Experimental procedure A between-subjects experimental design was used to compare the stepwise and gradual sugar reduction strategies. Children (6 –12 years old) were randomly allocated to two groups of similar size: stepwise sugar reduction (n = 74) and gradual sugar reduction (n = 73).

Participants were asked to attend 10 tasting sessions over an 11-week period. During the ﬁrst nine consecutive weeks, they evaluated the sugar-reduced grape nectar samples. Two weeks later, they performed a ﬁ nal liking test. A total of 30 participants did not attend all the sessions, giving as a result 66 children in the stepwise reduction group and 51 children in the gradual reduction group. No signi ﬁ cant di ﬀerences be- tween the two groups of children were found in their gender ( p= 0.82) and age ( p= 0.86). In addition, the groups did not signi ﬁcantly di ﬀer in their nectar consumption frequency ( p= 0.67) and stated nectar liking (p = 0.56), reported at the beginning of the test.

2.3.1. Sensory and hedonic perception of grape-nectars over a 9-week period Participants attended one session per week over a 9-week period.

Children in the stepwise sugar reduction strategy received the same nectar for three consecutive weeks. As shown in Table 1, they evaluated three nectars with di ﬀerent sugar concentration. Participants in the gradual sugar reduction strategy evaluated the sugar with the max- imum sugar concentration (10%) at the ﬁrst two weeks, and then re- ceived di ﬀerent nectar each week ( Table 1). At the ninth week both groups received the nectar with the lowest sugar concentration (4.3%).

In each session, they received a white plastic cup containing 100 mL of grape nectar. They were asked to drink as much as they wanted and to rate their overall liking using a 9-point hedonic scale (1 = super bad, 9 = super good) ( Kroll, 1990). They were also asked to answer a simple check-all-that-apply (CATA) questions composed of six sensory terms:

grape ﬂavor, very sweet, sweet, barely sweet, acid, and watery. Terms were selected by the researchers and a pilot test was conducted with children prior to the study to ensure understanding. The meaning of the six terms included in the CATA question was explained to the children in the beginning of the study.

2.3.2. Sensory and hedonic perception of grape-nectars two weeks after the 9-week period Two weeks after the 9-week period, an additional session was held, in which children evaluated the three nectar samples included in the stepwise sugar reduction strategy (added sugar concentration: 10.0%, 6.3% and 4.3%). The samples were evaluated by the two experimental groups following the same procedure described in the previous section.

Samples were presented according to an experimental design that was balanced for order and carry-over e ﬀects (Williams ’Latin Square de- sign).

2.4. Data analysis Analysis of variance (ANOVA) was used to analyze overall liking scores in both studies. Over the 9-week period, data were analyzed Table 1 Sugar concentration and overall liking of grape nectars served during a long- itudinal study with children, following two added sugar-reduction strategies:

stepwise and gradual.

Week Experimental Group Stepwise (n = 66) Gradual (n = 51) Added sugar concentration (g/ 100 mL) of the grape nectars Overall Liking Added sugar concentration (g/ 100 mL) of the grape nectars Overall Liking 1 10.0 8.2(1.2) a 10.0 8.0(1.7) a 2 10.0 7.9 (1.3) a 10.07.9(1.7) a 3 10.0 7.6(1.9) ab 9.07.6(1.8) ab 4 6.3 7.6(1.4) abc 7.87.6(2.0) ab 5 6.3 7.0(2.1) bcd 7.07.7(1.8) ab 6 6.3 7.4 (2.0) abc 6.37.3(2.3) ab 7 4.3 6.5(2.3) d 5.57.2(2.3) ab 8 4.3 6.8(2.2) cd 4.86.9(2.4) b 9 4.3 6.9(2.2) bcd 4.36.8(2.5) b Samples with diﬀerent lowercase letters in the same column are signi ﬁcantly di ﬀerent according to Tukey ’s test (p < 0.05). M. Lima et al. Food Quality and Preference 71 (2019) 163–167 164 considering experimental group, week and their interaction asﬁxed sources of variation. ANOVA was also used to analyze overall liking scores of each group, considering week as ﬁxed source of variation. In the test performed at the 11th week, experimental group, sample and their interaction were considered as ﬁxed sources of variation. Tukey ’s test was used for post-hoc pairwise comparisons at a signi ﬁcance level of 5%. Frequency of use of each CATA term was determined in both ex- periments by counting the number of children who used that term to describe each sample. Cochran ’s Q test followed by the sign test ( Meyners, Castura & Carr, 2013 ) were used to identify signiﬁcant dif- ferences among samples for each of the sensory terms for each ex- perimental group. In the 9-week period, a Generalized Linear Model (GLM) was used considering the use of the term (0/1) as dependent variable, and experimental group, week and their interaction as in- dependent variables. The 11th week data were analyzed using a GLM considering sample, experimental group and their interaction as in- dependent variables.

All data analyses were carried out using R software version 3.2.3 ( R Core Team, 2017 ).

3. Results 3.1. In ﬂuence of sugar reduction strategies on children’ s hedonic and sensory perception of grape nectars over a 9-week period 3.1.1. Hedonic perception Average overall liking scores ranged from 8.1 to 6.5 for the stepwise sugar reduction strategy and from 7.9 to 6.8 for the gradual sugar re- duction strategy ( Table 1). Children ’s overall liking scores were sig- ni ﬁcantly a ﬀected by week (F = 6.75, p < 0.001), but did not sig- ni ﬁcantly di ﬀer between groups (F = 1.04, p = 0.308) or the interaction between group and week (F = 0.87, p = 0.543).

As expected, overall liking scores signi ﬁcantly decreased with sugar reduction for both experimental groups. However, gradual sugar re- duction caused smaller changes in children ’s hedonic perception than the stepwise sugar reduction ( Table 1). For children in the stepwise strategy, the ﬁrst signi ﬁcant decrease in liking with respect to the control sample (10% sugar, week 1) occurred in the ﬁfth week (37% sugar reduction), whereas for children in the gradual reduction strategy overall liking signi ﬁcantly decreased in the eighth week (52% sugar reduction).

3.1.2. Sensory perception For both groups, signi ﬁcant di ﬀerences were found in the frequency of use of some of the terms of the CATA questions between the nectars tasted in di ﬀerent weeks. As shown in Table 2, children in the stepwise group increased their frequency of use of the terms watery(3rd week), acid taste (5th week) andbarely sweet (8th week), and decreased their frequency of use of the terms grapeﬂavor (6th week), very sweet(8th week) and sweet(7th week). Children in this group did not notice changes in the sensory characteristics of the nectars when sugar con- centration decreased from 10.0% to 6.3% (weeks 3 and 4). However, when the sugar content of the grapes was further reduced to 4.3% (weeks 6 and 7), they perceived changes in the sensory characteristics of the nectars, as evidenced by the decrease in the frequency of use of the term sweetand the increase in the frequency of use of the term watery .

For children in the gradual sugar reduction group smaller di ﬀer- ences in the frequency of use of the terms were found, which indicate that they perceived smaller di ﬀerences in the sensory characteristics of samples. This group of children increased their frequency of use of the terms watery (3rd week) and acid taste(6th to 8th weeks), and de- creased the frequency of use of the terms very sweet(7th week) com- pared to the ﬁrst sample.

The two groups did not largely di ﬀer in how they used the terms of the CATA questions to describe the samples ( Table 2). According to the Generalized Linear Model (GLM), signi ﬁcant di ﬀerences between the groups were found for the frequency of use of the terms sweet (p = 0.017), barely sweet(p < 0.001) and grapeﬂavor (p < 0.001), whereas the interaction between experimental group and week was not signi ﬁcant for all terms (p > 0.21). On average, children in the gradual reduction group used more frequently the terms sweet(51% vs. 44%) and grape ﬂavor (78% vs. 67%), and less frequently the term barely sweet than those in the stepwise reduction (13% vs 22%).

3.2. Sensory and hedonic perception of grape nectars with di ﬀerent sugar concentration evaluated after the 9-week period Overall liking scores were signi ﬁcantly a ﬀected by sample (F = 11.22, p < 0.001) and experimental group (F = 4.57, p = 0.033), whereas no signi ﬁcant interaction was found between sample and experimental group (F = 1.80, p = 0.166). As expected, the sample with the lowest added sugar content (4.3%) showed the lowest overall liking score for both experimental groups ( Table 3). At the ag- gregate level, children in the stepwise sugar reduction strategy gave signi ﬁcantly higher overall liking scores than those in the gradual sugar reduction strategy.

Diﬀ erences among samples were found in the frequency of use of the majority of the terms. As expected, the sample formulated with 4.3% of added sugar showed lower frequency of use of the terms very sweet andgrape ﬂavor intensity, and a higher frequency of use of the terms barely sweet ,acid taste andwatery than the control sample (10.0% added sugar). The two experimental groups did not signi ﬁcantly di ﬀer in how they used the terms of the CATA questions to describe the samples (p > 0.12). In addition, the interaction between experimental group and sample was not signi ﬁcant for all terms (p > 0.66).

4. Discussion and conclusions One of the main challenges of sugar reduction is maintaining con- sumers ’hedonic perception. This is particularly relevant in the case of children, who have a heightened preference for sweet taste ( Ventura & Menella, 2011). Sugar reduction should be unnoticeable for consumers, who slowly become accustomed to the sugar-reduced products. In the present work, the in ﬂuence of two sugar reduction strategies on chil- dren ’s sensory and hedonic perception were studied, using grape nectar as target product. Reducing the added sugar content of the grape nectars from 10.0% to 4.3% led to changes in the sensory characteristics perceived by children. In particular, a decrease in sweetness and an increase in acidity and watery was observed. The increase in the frequency of use of the term wateryfor describing sugar-reduced grape nectars can be explained by a reduction in grape ﬂavor intensity perception and viscosity, which can be related to both the reduction in sweetness and the increased concentration of water caused by the reduction of added sugar. These changes in the sensory characteristics of the nectars led to a decrease in overall liking. However, average overall liking scores were always higher than 6.5, which suggest that children had a positive hedonic attitude towards the sugar-reduced nectars. Despite the re- duction of 57% in the added sugar concentration of grape nectars commercialized in Brazil (from 10.0% to 4.3%) caused a signi ﬁcant decrease in the overall liking, the average score still indicated a positive hedonic attitude towards the product. This stresses the feasibility of sugar reduction to decrease sugar intake at the population level.

Although there were no major di ﬀerences between the gradual and stepwise reduction, results showed that the gradual reduction strategy led to smaller changes in children ’s sensory and hedonic perception than the stepwise strategy. Therefore, gradual reduction could be re- garded as more e ﬀective for maintaining children ’s satisfaction with the product. Food companies should not be concerned about children dis- liking their product and switch to the competitors if gradual sugar M. Lima et al. Food Quality and Preference 71 (2019) 163–167 165 reduction programs based on diﬀerence thresholds are implemented, as they would not perceive the di ﬀerence. According to results from pre- vious studies conducted with children and adults, di ﬀerence thresholds for added sugar in beverages correspond to 6.2 –8.5% ( Lima et al., 2018; Oliveira et al., 2016; Hoppert, Zahn, Puschmann, Ullmann, & Rohm, 2012; Pineli et al., 2016 ), which would correspond to the sugar re- duction of each step.

The advantages of gradual reformulation programs have been ex- tensively explored for salt. In this sense, Bobowski, Rendahl, and Vickers (2015) reported that gradual salt reduction in tomato juice was more e ﬀective than an abrupt reduction for maintaining consumers liking throughout 16-week period. Gradual salt reduction programs have proven to be successfulat both the commercial ( Bratt & Tamman, 2010 ) and the population levels ( He, Brinsden & MacGregor, 2014; Public Health England, 2017 ).

Two weeks after the longitudinal study, children in the stepwise sugar reduction strategy showed higher liking for the nectars than children in the gradual sugar reduction strategy. This di ﬀerence can be explained considering di ﬀerences in their expectations. Children in the stepwise reduction strategy were more times exposed to the nectar with the lowest sugar content (4.3%) than those in the gradual reduction and, therefore, expected to receive nectars with low sugar. However, when they received nectars with higher sugar content, their expecta- tions were discon ﬁrmed and they increased their liking. It is also pos- sible to suggest that the learning process played a role. In this sense, the overall liking of the nectar sample with the lowest added sugar content signi ﬁcantly increased for the stepwise group after the 9th week period compared to the ﬁrst time they taste it. This suggests that exposure to this sample over a 3-week period led to an increase in liking for the sample with the lowest sugar content. Several studies have consistently demonstrated that liking for foods increases with repeated exposure, for both adults and children ( Cooke, 2007). Considering that children were only exposed to the nectars once a week, a 3-week waiting period be- tween gradual sugar reduction steps seems reasonable. However, it should be stressed that the 3-week waiting time im- plemented in the stepwise reduction was not enough to modify chil- dren ’s preference for sweet taste in grape nectar. Similar results have been reported by Biguzzi, Lange and Schlich (2015) when studying changes in liking after exposure to sugar-reduced cookies over ﬁve testing sessions conducted once a week with adult consumers. Further research in this respect should be carried out for providing re- commendations for the design of gradual sugar reduction programs. In addition, di ﬀerent results could have achieved whether the children have tried the nectars more than once a week. Therefore, this limitation has to be taken into account and in future studies participants have to try the product more times, as reported by Bobowski et al. (2015).

Acknowledgements The authors are indebted to Conselho Nacional de Desenvolvimento Cientí ﬁco e Tecnológico (CNPq, Brazil) (CNPq: 401470/2014-8) and Espacio interdisciplinario (Universidad de la República, Uruguay) for ﬁ nancial support. The authors also thank the Escola Técnica do Rio de Janeiro (ETERJ) for the children participation.

Table 2 Percentage of children who used each of the terms of the CATA question in the 9-week experiment in the stepwise and gradual added sugar reduction scheme.

Experimental group Week Added sugar concentration Acid taste Very sweet Sweet Barely sweet Grape ﬂavor Watery Stepwise 1 10.0% 2 c 13a 34a 6b 51a 3c 2 10.0% 4 abc 16a 32a 9b 51a 8bc 310.0% 6 abc 8ab 35a 10b 48a 11b 46.3% 7 abc 14a 27ab 14ab 46a 10b 56.3% 9 ab 13a 27ab 19ab 44a 15b 66.3% 8 ab 8a 32a 12ab 40bc 16b 74.3% 10 a 5abc 22b 21ab 34bc 35a 84.3% 10 a 2cd 25ab 23a 40bc 31a 94.3% 9 a 9ab 25ab 17ab 43abc 27a Gradual110.0% 1 b 7b 28ab 5ab 42a 3c 210.0% 4 b 12a 30a 3b 42a 4bc 39.0% 5 ab 14a 28ab 3b 39a 8b 47.8% 5 ab 9b 22ab 3b 41a 10b 57.0% 6 ab 5bc 27ab 4b 39a 10b 66.3% 9 a 8b 20b 6ab 40a 18a 75.5% 8 a 1c 28ab 11a 39a 17a 84.8% 8 a 1c 25ab 12a 39a 19a 94.3% 6 ab 4bc 26ab 12a 39a 21a The frequency of use of terms with di ﬀerent lowercase letters within a column and experimental group are signi ﬁcantly di ﬀerent between weeks according to the sign test (p < 0.05).

Table 3 Overall liking and percentage of children who used each of the terms of the CATA questions for the grape samples with di ﬀerent added sugar concentration (% of nectar weight) submitted to the stepwise and gradual sugar reduction scheme two weeks after the 9-week period.

Group Added sugar concentration Overall liking Acid taste Very sweet Sweet Barely sweet Grape ﬂavor Watery Stepwise 10.0% 8.2(1.1) a 2b 26a 34a 1c 53a 1c 6.3%7.8(1.5) a 5a 7b 41a 7b 54a 7b 4.3%7.3 (1.8) b 14a 3b 27a 18a 40b 22a Gradual 10.0% 7.6 (2.0) a 1b 20a 25a 1b 42a 1c 6.3%7.9 (1.7) a 5ab 4b 25a 5a 41a 7b 4.3%6.5 (2.6) b 12a 1b 15b 17a 31b 21a Overall liking scores with diﬀerent lowercase letters within an experimental group are signi ﬁcantly di ﬀerent between added sugar concentrations according to Tukey ’s test (p < 0.05). The frequency of use of terms within a column and experimental group with di ﬀerent letters is signi ﬁcantly di ﬀerent according to the sign test (p < 0.05).

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